Re-use of electroporation cuvettes?
Mr R D Williams
after washing out the transformation mix with SOC? - i.e. Is there a cleaning
protocol that effectively removes traces of both salt and DNA from the
cuvette? Is simple washing with distilled water sufficient? How many times is
it safe to re-use these expensive little things before arcing is likely?
Thanks in advance,
Richard.
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Richard Williams ls...@csv.warwick.ac.uk
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John Gosink
(Mr R D Williams) wrote:
> Does anyone have any experience of re-using Bio-Rad's electroporation cuvettes
< snip >
> it safe to re-use these expensive little things before arcing is likely?
There were several posts on this subject about half a year ago. I
think the upshot was something like soak the cuvettes with acid or bleach
for a bit to destroy any DNA that was there, then rinse the heck out of
them with dH2O to get rid of any salt.
Personally, I have been using the following procedure:
1) Rinse them out with 70% ethanol (to kill the bacteria and get most
of the
gunk physically out of the cuvette)
2) Soak them in 0.1 M Nitric Acid for about 10 minutes
3) Rinse them 10 times with STRONG squirts of dH2O to reach down into
the
very bottom of the cuvette
4) Rinse them 2 times with 95% ethanol and let air dry
I'm going on the 4th and 5th reuse of the same cuvettes and haven't had any
noticable problems with sparcing or "positives" in my "negative" controls.
Good luck.
-John
Manuel G. CLAROS
wrote:
>
>I don't see why others are having problems. I've been using the
>SAME few cuvettes for 12 MONTHS now. (roughly 60 "zaps" each).
I also have been using them for a year. I simply clean them well with dH2O after
use, and then wash intensively with ethanol.
Manuel G.
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Przemko
>Does anyone have any experience of re-using Bio-Rad's electroporation cuvettes
>protocol that effectively removes traces of both salt and DNA from the
>cuvette? Is simple washing with distilled water sufficient? How many times is
>
>
>Richard.
>--
>-----------------------------------------------------------------------------
>Richard Williams ls...@csv.warwick.ac.uk
>-----------------------------------------------------------------------------
Hi!
No problem. We reuse them many times over. The protocol is very simple.
ASAP after electroporations wash with tap water (I put a blue tip on the
end of the rubber tubing I have on my faucet so I can get rather high
water pressure), wash with deionized water, wash with 70% ethanol,
dry on paper towels (upside down) and reuse.
NO AUTOCLAVING, NO BLEACH otherwise they will flash after two or three
attempts to reuse. Also if a cuvet flashes then it goes IMMEDIATELY
into garbage.
Hope it helps
Przemko
Harry Menegay
>Does anyone have any experience of re-using Bio-Rad's electroporation cuvettes
>after washing out the transformation mix with SOC? - i.e. Is there a cleaning
>protocol that effectively removes traces of both salt and DNA from the
>cuvette? Is simple washing with distilled water sufficient? How many times is
>it safe to re-use these expensive little things before arcing is likely?
I don't see why others are having problems. I've been using the
SAME few cuvettes for 12 MONTHS now. (roughly 60 "zaps" each). I simply
rinse well with dd H2O, and UV the suckers for a few minutes in our UV
stratalinker. I'm about to change to a fresh set just because I'm worried
what all the UV may be doing to the plastic. The UV kills all the bugs and
trashes any old vector as well. With a good batch of electrocompetent cells
and my DNA resuspended in H2O (no ligation salt) I still get time constents
of 4.0 - 5.0. One of them did arc on me after a few uses, but that apparently
was due to too much salt in the ligation. I've also never had any problems
of contamination from old vector, due I'm sure to the UV treatment.
Good Luck,
Harry
mal...@sara.cc.utu.fi
> No problem. We reuse them many times over. The protocol is very simple.
> ASAP after electroporations wash with tap water (I put a blue tip on the
> end of the rubber tubing I have on my faucet so I can get rather high
> water pressure), wash with deionized water, wash with 70% ethanol,
> dry on paper towels (upside down) and reuse.
> NO AUTOCLAVING, NO BLEACH otherwise they will flash after two or three
> attempts to reuse. Also if a cuvet flashes then it goes IMMEDIATELY
> into garbage.
This is not quite true, because I've been using cuvettes over 20 times now and
after every time I have soaked cuvettes in diluted bleach for 20 min. After
that cuvettes have been rinsed thoroughly in dH2O, then rinsed with 70 % EtOH
and finally with 100 % EtOH. Then I inverted cuvettes onto a clean paper towel
in flaminar food and let them dry completely. After that they are ready to use
again. And very RARELY I have seen any sparking at all. Whether it is sparking
or not is more up to your sample and the amount of salts in that.
Marko
Marko Laine
Univ. of Turku Mal...@sara.cc.utu.fi
Dept. of Plant Physiology Mal...@polaris.utu.fi
BioCity 6th floor \\ // Fax: +358-21-633 8075
20520 Turku (-o-o-) Tel: +358-21-633 8076
Finland ----oOOo----(_)----oOOo---- NMT: 949-864 954
- Dr. Livingstone, I presume...
Peter M. Muriana
>> Hi!
>> No problem. We reuse them many times
>> NO AUTOCLAVING, NO BLEACH otherwise they will flash after two or three
>> attempts to reuse. Also if a cuvet flashes then it goes IMMEDIATELY
>> into garbage.
>
>This is not quite true, because I've been using cuvettes over 20 times now and
>after every time I have soaked cuvettes in diluted bleach for 20 min. After
>that cuvettes have been rinsed thoroughly in dH2O, then rinsed with 70 % EtOH
> I have seen any sparking at all. Whether it is sparking
>or not is more up to your sample and the amount of salts in that.
>
>Marko
>
>
>Marko Laine
>Univ. of Turku Mal...@sara.cc.utu.fi
>Dept. of Plant Physiology
Marko, I presume from your affiliation you may be electroporating cells
other than bacteria which often require high voltages compared to other
types of cells; the arcing (in our hands) also depends on what type of
parallel resistance is chosen on the electroporation device.
Regards, Peter
>Finland
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